Team:WashU/Week4

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Revision as of 16:38, 22 June 2012

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YLC

The team has finished up the PowerPoint to be presented on the day of the YLC Outreach. In addition, the team made plates as demos for the children using CFP, YFP, GFP and RFP. [Pictures to follow.]

To test whether we had the proper constructs for our different fluorescent proteins, the team ran a gel of the digests, shown below.

Synechocystis

We plated a Kanamycin resistant synechocystis on plates made with noble agar and glucose, noble agar and no glucose, bacteriological agar and glucose, and bacteriological agar and no glucose.


Result of agar test after four days. Notation: N=noble agar, B=bacteriological agar, -G=without glucose (autotrophic) and +G=with glucose(mixotrophic)

E. coli

The transformations from last week seemed to have worked. The CFP is very very light and looks almost like GFP. We think it may be because our UV light isn't very strong.

The team has continued to attempt to biobrick together the necessary genes that create the carotenoid pathway in E. coli (see last week for more information). To do this, the team has decided to follow the biobrick assembly protocol once more, synthesizing the upstream, downstream and plasmid parts, and then connect them. In order to increase the efficiency and yield of this assembly, gel purification was used to isolate the plasmid for the biobrick assembly. Below is the gel with the upstream, downstream and plasmid (The experiment was done twice.) potential%2520product4.jpg

However, instead of using the old promoter J23100, the team has decided to instead use part BBa_J23119, the strongest promoter of that constitutive promoter family.

Modeling

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 The transformations from last week seemed to have worked. The CFP is very very light and looks almost like GFP. We think it may be because our UV light isn't very strong.
-The team has continued to attempt to biobrick together the necessary genes that create the carotenoid pathway in E. coli (see last week for more information). To do this, the team has decided to follow the biobrick assembly protocol once more, synthesizing the upstream, downstream and plasmid parts, and then connect them. However, instead of using the old promoter J23100, the team has decided to instead use part BBa_J23119, the strongest promoter of that constitutive promoter family.
+The team has continued to attempt to biobrick together the necessary genes that create the carotenoid pathway in E. coli (see last week for more information). To do this, the team has decided to follow the biobrick assembly protocol once more, synthesizing the upstream, downstream and plasmid parts, and then connect them. In order to increase the efficiency and yield of this assembly, gel purification was used to isolate the plasmid for the biobrick assembly. Below is the gel with the upstream, downstream and plasmid (The experiment was done twice.)
+https://lh6.googleusercontent.com/-c5PpgOPE9GE/T-Serah0tHI/AAAAAAAAAJI/c-YW88UISPM/s800/potential%2520product4.jpg
+However, instead of using the old promoter J23100, the team has decided to instead use part BBa_J23119, the strongest promoter of that constitutive promoter family.
 ==Modeling==